Carotenoids are coloured Carotenoids are coloured pigments that found in nature which have their specific colour range of yellow, orange and red colours. Colour plays an important role to make sure the products are attractive to the consumers. The natural colorant is colouring that can be obtained from natural sources such as plants, vegetables, animals and minerals. This study established that the working scheme via open column chromatography (OCC) established from pumpkin and sweet potato demonstrated that isolation of the β-carotene and lutein pigment were successful with 97.2% and 95.46% purity with the coefficient of correlation of 0.9961 and 0.9959 respectively. The β-carotene extract has presented strong activity on DPPH scavenging radicals with the percentage of inhibition at 99.45% in 0.008 mg/ml concentration as compared to lutein, which contributed 65.6% at 0.5mg/ml. As for antimicrobial activity, a positive result for both β-carotene and lutein pigment for bacterial and fungal species except for Tricho sp. In this study, the results from chromaticity stability of PLA-β-carotene and lutein coating from 1mg/ml up to 3mg/ml treated with different pH value from 3.0 to 11.0, as well as the time of exposure from day 1 until day 15 revealed that there is no stable relationship between pH, yellow colour concentration and period of time in response to light. Marked differences were observed for PLA-β-carotene coating treated with different concentration of NaCl for 1 and 2 mg/ml concentration of carotenoid pigment for a period of time of 10 days and PLA-lutein coating treated with different temperature from -25°C to 50°C when exposed for 6 hours. In term of light exposure, PLA-β-carotene coating with lower concentration and PLA-lutein with higher concentration were found to slow the degradation process. In conclusion, this data suggests that although different carotenoid pigment concentrations had influenced chromaticity stability, light and time also affected pigment colour difference.

Nowadays, halal products are gaining wider recognition as a new benchmark for safety and quality assurance. As a consequence the commercial development of microalgae are established due to their high value chemicals, for examples, β-carotene, astaxanthin, phycobilin pigments and algal extracts for cosmaceuticals products. Therefore, many researchers have gained interest to study the potential of microalgae as new valuable chemicals and other products sources. The aim of the research is to explore new sources of pigments to be used as halal food colorants. This quest is not only directed in finding natural alternatives for synthetic dyes, but also to discover new taxons for the carotenoid production. Thus, there is a solid need to investigate the potential of natural pigments particularly carotenoids in microalgae to be fully utilised and commersialised especially in halal market, health advantages, food products and dye technology. A total of 10 species were evaluated for quantitative and qualitative carotenoid composition namely Chlorella fusca, Chlorella vulgaris, Selenastrum capricornutum, Pandorina morum, Butyriciococcus sudeticus ,Chlorococcum, Pseudanabaena, Scenedesmus, Ankistrodesmus, and Butyrocicoccus Brauni.Chlorella fusca (chlorophyta) and Pseudanabaena(cyanobacteria) and found three types of carotenoid out of them which are β-carotene, β-carotene and lutein. Species that was having all three types of carotenoid was Pandorina morum (Figure 4.8) with lutein (54.64±3.11 mg/g DW) , Beta cryptoxanthin (2.38±0.56 mg/g DW) and Beta carotene (1.39±0.38 mg/g DW) Manipulation of nutrient and nitrate concentration, manipulation of carbon sources. Two types of microalgae were choosen represent their division which are Chlorella fusca (Chlorophyta) and Pseudanabena (Cyanobacteria) were subjected to nutrient and manipulation stress studies. There were cultured using Bold’s Basal Medium (BBM), Bristol medium and Bold’s Basal Medium (BBM) modified with vitamin B with different nutrient and carbon sources. There were three types of carotenoid had produced which are zeaxanthin (21248.7 ± 240.61mg/g DW), violaxanthin (16299±423.09mg/g DW) and lutein (2335.53±69.82mg/g DW) with their highest number respectively. Nitrate concentration also been manipulated in all types of medium that been used and it had produced lutein. The highest lutein number was produced in cultivation of Pseudanabaena in Bristol medium with 0.5 fold nitrate concentration (1317.91±40.13mg/g DW). Chlorella fusca and Pseudanabena also been cultured in BBM with the addition of different elicitors which are salicylic acid (SA), polyethylene glycol (PEG) and sodium chloride (NaCl). All of these elicitors had initiate production of lutein. Among of those three elicitors, this studies had found that the implication of 25mg/L of SA had yield highest lutein concentration (666.45±24.66mg/g DW).Cell concentration of microalgae also been recorded for all studies except for profiling. Cell count were done using haemocytometer under microscope and growth pattern of microalgae been plotted to monitor microalgae cell density